Stem cell transfection - (Dec/15/2005 )

Hi!

We are using electroporation to transfect mouse ES cells and our transfection efficiency is really low. Has anyone tried viral transfection in ES cells?

Thanks a lot!

-wakeupjune-

I plan on doing primary and breast stem cell transfections. Due to them being hard to transfect we will have to use viral transfection and our neighbouring lab uses lenti-virus for that matter. Before doing so we will test whether our cationic lipid transfectants can achieve this as well, as it will be much more cost effective.Anyway, try at the regular companies for info on viral delivery - Dharmacon, Invitrogen etc.

-avestin-

My labmate has tried several transfection reagents on mouse ES cell line including lipo2000, fugene-6, Superfect, TransIT, Gencarrier-2, GenePORTER-2......among these, gencarrier-2 showed much better efficiency (~30-40%) and lowest toxicity. She later tried lentiviral system also, but again, she need to add some sort of agents (DEAE-dextran) to boost viral infection efficiency and need to concentrate virus to get higher titer.